1. We created fusion chimeras between GR and other nuclear receptors that manifest ligand dependent cytoplasmic/nuclear translocation for the chimeric receptor in response to the heterologous ligand. The approach has been successfully developed for the glucocorticoid receptor (GR), the retinoic acid receptor (RAR), the estrogen receptor (ER), the aryl hydrocarbon receptor (AhR), the progesterone receptor (PR), and the thyroid hormone receptor (ThR). We developed cell lines which express fluorescently tagged versions of the receptor chimeras. We have utilized this fluorescent protein fusion technology to show that U.S. water supplies are heavily contaminated with chemicals that have glucocorticoid and androgen activity. US patent No. 20150168416 was granted covering licensing of this technology. We have collaborated with multiple intramural and extramural investigators to track hormone contamination in U.S. waterways, and explore the implication of these findings for human health. 2. Transcription factors dynamically bind to chromatin and are essential for the regulation of genes. While a large percentage of these proteins appear to self-associate to form dimers or higher-order oligomers, the stoichiometry of DNAbound transcription factors has been poorly characterized in vivo. The glucocorticoid receptor (GR) is a ligand regulated transcription factor and its oligomerization state has been implicated in clinical outcome of therapy. Although widely believed to act as a dimer, we have shown that GR is predominantly a tetramer when bound to its target DNA. Using a unique set of imaging techniques coupled with a cell line containing an array of DNA binding elements, we found that DNA binding triggers an inter-domain allosteric regulation of the GR, followed by a change in its oligomeric state. These findings demonstrate a new step in steroid receptor activation an open new doors to the rational design of novel GR ligands. We discovered that a mutation in the GR DBD causes the receptor to constitutively adopt a tetrameric form of the receptor. When expressed in GR knock out cells, all nuclear receptor is in the tetrameric state. We have begun an exploration of the genome wide behavior for this receptor state.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIABC010308-20
Application #
9779611
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
20
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
Zip Code
Lynch, Caitlin; Sakamuru, Srilatha; Huang, Ruili et al. (2017) Identifying environmental chemicals as agonists of the androgen receptor by using a quantitative high-throughput screening platform. Toxicology 385:48-58
Presman, Diego M; Hager, Gordon L (2017) More than meets the dimer: What is the quaternary structure of the glucocorticoid receptor? Transcription 8:32-39
Stavreva, Diana A; Varticovski, Lyuba; Levkova, Ludmila et al. (2016) Novel cell-based assay for detection of thyroid receptor beta-interacting environmental contaminants. Toxicology 368-369:69-79
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Stavreva, Diana A; George, Anuja A; Klausmeyer, Paul et al. (2012) Prevalent glucocorticoid and androgen activity in US water sources. Sci Rep 2:937
Biddie, Simon C; John, Sam; Sabo, Pete J et al. (2011) Transcription factor AP1 potentiates chromatin accessibility and glucocorticoid receptor binding. Mol Cell 43:145-55
Sung, Myong-Hee; McNally, James G (2011) Live cell imaging and systems biology. Wiley Interdiscip Rev Syst Biol Med 3:167-82

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